OBJECTIVETo study the role of integrin alpha 6 in cell attachment, spread, survival/proliferation and differentiation of human hepatocellular carcinoma (HCC) BEL-7402 cells on various substrates by monoclonal antibody against the extracellular domain of alpha 6 subunit (IA6ED McAb).

METHODSThe effect of McAb on attachment and spread of BEL-7402 cells on LN or FN substrate was examined. MTT analysis was used to examine the cell survival/proliferation, gelatin zymography to the matrix metalloproteinases (MMPs) secreted by BEL-7402 cells, and microparticle immunoabsorbent kit to AFP secretion while the cells were cultured on LN-, FN- or Matrigel-coated substrates.

RESULTSThe cell attachment, spread and survival/proliferation were inhibited. Moreover importantly, the malignant cell dedifferentiation and abnormal differentiation on LN-coated substrate were also strongly inhibited by IA6ED McAb.

CONCLUSIONLN and integrin alpha 6 regulate human HCC cell phenotypes of survival/proliferation and differentiation. The phenotypes of dedifferentiation and abnormal differentiation can be reversed by blocking the interaction between integrin alpha 6 and LN using IA6ED McAb, which may lower metastatic potency of tumor cells.

Antibodies, Monoclonal ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; Cell Adhesion ; drug effects ; Humans ; Integrin alpha6 ; immunology ; physiology ; Laminin ; physiology ; Liver Neoplasms ; pathology ; Phenotype ; Receptors, Laminin ; physiology ; Tumor Cells, Cultured

@@

We designed specific primers and fluorescence-labeled probes to develop real-time and conventional RT-PCR assays for detection of human coronavirus NL63 or HKU1. Subsequently, experiments were undertaken to assess diagnostic criteria such as specificity, sensitivity and reproducibility. The detection limit of the real-time RT-PCR assays was 10 RNA copies per reaction mixture. No cross-reactivity was observed between RNA samples derived from designed HCoV and other HCoV or human metapneumovirus. A total of 158 nasopharyngeal swab specimens collected from adult patients with acute respiratory tract infection in Beijing were screened for the presence of human coronavirus NL63 and HKU1 by using real-time RT-PCR and conventional RT-PCR method. The fluorescence quantitative RT-PCR method detected six specimens positive for human coronavirus NL63, five specimens positive for human coronavirus HKU1; and conventional RT-PCR method detected three HCoV-NL63 positive and three HCoV-HKU1 positive, respectively. The convention RT-PCR products of positive samples were obtained and sequence analysis confirmed the reliability of the above methods. In summary, the real-time RT-PCR assay for HCoV- NL63 or HKU1 was more sensitive than conventional RT-PCR and with less time (less than 4 hours) for completion. It may be suitable for molecular epidemiological surveillance and clinical diagnosis for human coronavirus NL63 and HKU1.
Coronavirus ; classification ; genetics ; isolation & purification ; Humans ; Nasopharynx ; virology ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

OBJECTIVETo know the etiology, prevalence, clinical symptoms associated with the infection of the HCoV-229E in the respiratory specimens sampled from adult patients in Beijing.

METHODS158 nasopharyngeal swab specimens were collected from adult patients with fever in Beijing between October and December, 2007. We performed the screening of HCoV-229E by real-time RT-PCR and sequencing of HCoV-229E gene fragments derived from conventional PCR. At meantime, we also screened the HCoV-229E positive samples for the co-infection with HCoV-NL63, HCoV-HKU1 and HMPV by real-time RT-PCR. Finally, demographic and clinical data associated with HCoV-229E infection were examined retrospectively.

RESULTSWe detected 103 (62.5%) of 158 specimens were positive for HCoV-229E by real-time RT-PCR. When tested for other respiratory viruses, 26 HCoV-229E positive patients were found to be co-infected with other viruses. Of which HCoV-NL63 was observed in 3 specimens (11.5%), HCoV-HKU1 in 3 (11.5%) and HMPV in 20 (76.9%). The main clinical manifestations were noted as: headache (in 70.9%), sore throat (69%), chills (68%), cough (33%), sputum (21.3%), rhinorrhea (21.4%), nasal obstruction (16.5%), and a few of patients were visible as vomiting (6.8%), dyspnea (3.9%), diarrhea (in 1.9%). The rate of HCoV-229E infection in adult patients was found no relative with age and gender.

CONCLUSIONOur data showed that HCoV-229E is a common and important pathogen in adult patients with acute respiratory symptoms but usually resulted in milder influenza-like illnesses. There might have a local outbreak of HCoV-229E infection in Beijing, Oct-Dec, 2007.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Coronavirus 229E, Human ; classification ; genetics ; isolation & purification ; Coronavirus Infections ; diagnosis ; epidemiology ; virology ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Phylogeny ; Respiratory Tract Infections ; diagnosis ; epidemiology ; virology ; Retrospective Studies ; Young Adult

OBJECTIVETo develop and optimize the molecular detection assays for recently identified human coronavirus (HCoV) infection.

METHODSBased on the 208 base pair(bp) sequence of novel HCoV reported by HPA of UK, we designed and obtained several pairs of primer (F-1, R-1; F-2, R-2) and Taqman probes (TZ1,TZ2) for detection of novel HCoV. Two of probes were modified with LNA (LNA-TZ1, LNA-TZ2). Then, RT-PCR and various real time RT-PCR assays were developed and optimized in this study. We also compared our assays with the real time RT-PCR assays reported recently by Europe team based on upE or ORF1b target.

RESULTSThe RT-PCR or real time RT-PCR assays for novel HCoV were developed without cross-reactivity with other HCoV and several common respiratory viruses using clinical specimen panel. The analytical sensitivity of assays were less than 50-500 copies per reaction and the detection was improved when Taqman probe modified with LNA-tagged, compared to no LNA-tagged in real time RT-PCR assays. The upE and LNA-TZ1 based assays were better than others.

CONCLUSIONThe molecular detection sensitivity and specificity of TaqMan-based real time PCR assay could be improved when probe tagged with LNA. The upE or LNA-TZ1 based real time RT-PCR assay was recommend for detection of novel HCoV. This study laid a foundation for improving the performance of novel HCoV detection.

Coronavirus ; classification ; genetics ; isolation & purification ; DNA Primers ; genetics ; Humans ; Oligonucleotides ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To analyze the implementation of intervention programs targeted on AIDS high risk sexual transmission groups since 2008,when the relative prevention and control information systems on HIV/AIDS were developed.Methods Data from both aggregated interventions and sentinel surveillance programs from 2008 to the end of 2014 were used.Descriptive statistics were performed to analyze the trends of implementation on high risk groups including men who have sex with men,female sex workers (FSW) and migrant workers.Results From 2008 to 2012,the monthly average numbers receiving intervention programs and the average monthly coverage rate on intervention for MSM,increased from 49000 to 252000,and from 8.6％ to 78.5％ respectively.The FSW related indicators increased from 329 000 to 625 000,and from 30.9％ to 87.0％ respectively.Above indexes on the two populations had dropped slightly in 2013 and 2014.Sentinel surveillance data showed that knowledge and behavior indicators observed from the MSM and FSW populations increased annually.The coverage of intervention programs on migrant workers increased from 4.7％ to almost 10.0％,but the surveillance data on migrant men showed that the knowledge and behavior indicators were still lower than the other high-risk groups.Conclusion Intervention related to sexual transmission on HIV/AIDS among high-risk populations were effectively implemented,with some achievements seen.However,as sexual contact currently became the main route of AIDS epidemic,new challenges called for serious attention.

OBJECTIVE: LAPTM4B-35 protein is one of the isoforms that are encoded by a cancer driver gene, LAPTM4B. This gene was primarily found and identified in our lab of Peking University School of Basic Medical Sciences. The LAPTM4B-35 protein and its encoded mRNA are significantly over-expressed in a variety of cancers, such as hepatocellular carcinoma (HCC), lung cancers (including non small-cell lung cancer and small-cell lung cancer), stomach cancer, colorectal carcinoma, pancreatic cancer, gallbladder cancer, cholangiocarcinoma, breast cancer, prostate cancer, ovarian cancer, cervical cancer, endometrial cancer, and so on. It has firmly demonstrated through lab experiments either in vivo or in vitro, as well as clinical studies that the over-expression of LAPTM4B-35 can promote cancer growth, metastasis, and multidrug resistance. Specially, the expressive level of LAPTM4B-35 is associa-ted with recurrence of HCC. The aim of this study is to identify the release of LAPTM4B-35 protein from hepatocellular carcinoma into blood of HCC patients and into the medium of cultured HCC cells, and to identify its possible form of LAPTM4B-35 protein existed in blood and cell culture medium, as well as to explore the possibility of LAPTM4B-35 protein as a novel HCC biomarker for diagnosis of HCC and prognosis of HCC patients. METHODS: Immunobloting (Western blot) and enzyme-linked immunosorbent assay (ELISA) were used for identification of LAPTM4B-35 protein in the blood of HCC patients and normal individuals. Ultrafiltration and ultracentrifugation were used to isolate and purify exosomes from the culture medium of HCC cells. RESULTS: LAPTM4B-35 protein existed in the blood from HCC patients and normal donors that were demonstrated through Western blot and ELISA. LAPTM4B-35 was also released into the culture medium of HCC cells in the form of exosomes. Preliminary experiments showed that the average and the median of LAPTM4B-35 protein level in the blood of HCC patients (n=43) were both significantly higher than that in the blood of normal donors (n=33) through sandwich ELISA. CONCLUSION It is promising that the LAPTM4B-35 protein which is released from HCC cells in the form of exosomes into their extraenvironment may be exploited as a novel cancer biomarker for HCC serological diagnosis.
Biomarkers, Tumor ; Carcinoma, Hepatocellular/diagnosis* ; Humans ; Liver Neoplasms/diagnosis* ; Male ; Membrane Proteins/genetics* ; Oncogene Proteins ; Prognosis

Objective:To evaluate the enrollment rate, mutation rate and causes of variability the clinical pathway of bronchopneumonia.Methods:The enrollment rate, completion rate, variation and reasons of the clinical pathway in Beijing Children′s Hospital, Capital Medical University from January 2012 to December 2016 were retrospectively collected.Data of patients after the clinical pathway of bronchopneumonia in other tertiary class A hospitals were gathered by questionnaires, and the enrollment rate, completion rate, variation rate and reasons were analyzed.Results:(1)At the end of 2016, 11 of the 13 hospitals included in this study had implemented the clinical pathway for 5 years, 1 hospital for 3 years, and 1 hospital for 2 years.(2) Eleven hospitals provided their enrollment rates.The enrollement rate of 2 hospitals was<50%, and that of 9 hospitals was>80%.The annual completion rate of Beijing Children′s Hospital was ≥75%, and the completion rates offered by 8 hospitals were basically >70%.(3) Since the implementation of the clinical pathway for 5 years in Beijing Children′s Hospital, a total of 427 cases were enrolled of which 93 cases were mutated (variability 21.78%). The variability of 5 hospitals was maintained at <15%.The variability of 3 hospitals decreased with the implementation years, and became qualified.The variability of 1 hospital first rebounded and then controlled; 1 hospital increased by 27.65%; 1 hospital was first controlled and rebounded; 1 hospital was always >15%.The main cause of the mutation was coexisting diseases, complications, progression of the disease, or correction of the first diagnosis, etc.Conclusions:The completion rate of tertiary class A hospitals meets the requirements of national policy.However, the enrollment rate needs to be improved, and the variation rate among different hospitals differs a lot.Further implementation of the clinical pathway should be strengthened and monitored.

Objective: To investigate the changes of proportion on both consistent condom use and syphilis infection among low-fee female sex workers aged 35 and above (LFSW), in order to provide evidence for targeted intervention strategies. Methods: A total of six cities-Liuzhou city and Pingnan couty of Guigang city of Guangxi Zhuang Autonomous Region, Jinghong city of Xishuangbanna Dai autonomous prefecture and Dali city of Dali Bai autonomous prefecture of Yunnan province, Zhangjiajie city and Jianghua Yao autonomous county of Yongzhou city Hunan province were involved in this study, with 60 eligible participants needed in each city, estimated through a pre-study. The first cross-sectional survey was completed from October 2012 to January 2013. Face-to-face questionnaire interview was carried out to collect information on socio-demography, work-related information and condom use situation. Blood was collected for syphilis testing. The second cross-sectional survey was carried out from June to September, 2015 under the same procedure. Results: A total of 371 and 403 eligible participants were included in the first and second survey, respectively. When comparing the two surveys, we noticed that the average age showed a slight change, from 42.4 years to 43.8 years old (t=3.537, P<0.001) and the average price for every commercial sex exchange increased from 36.8 RMB to 49.5 RMB (t=11.961, P<0.001). In the first survey, 46.9% (174/371) of the participants had more than two years of experience working as LFSW, compared to 61.3% (247/403) in the second survey (χ(2)=16.125, P<0.001). Also, 46.9% (174/371) of the participants consistently used condoms with clients in the past month in the first survey versus 64.3% (259/403) (χ(2)=23.641, P<0.001) in the second one. Rates of syphilis infection were found from 15.9% (59/371) in the first survey reduced to 7.2% (29/403) in the second survey (χ(2)=14.533, P<0.001). Conclusion: Compared with the first survey, the proportion of consistent condoms use showed an increase. Although the proportion of syphilis infection decreased in the second survey, the scope did not meet the criteria on syphilis, set by the government. Targeted intervention strategies on condom promotion and syphilis control should be implemented consistently in this population.
Adult ; China/epidemiology* ; Condoms/trends* ; Cross-Sectional Studies ; Female ; Humans ; Prevalence ; Risk Factors ; Safe Sex ; Sex Work ; Sex Workers ; Sexually Transmitted Diseases/epidemiology* ; Surveys and Questionnaires ; Syphilis/prevention & control* ; Syphilis Serodiagnosis

【Objective】 To explore the attention and its relationship with anxiety in patients with glaucoma.【Methods】A prospective case-control study was done on 20 primary open angle glaucoma（POAG）patients and 28 normal subjects. Response time with and without disturbance was examined by using Matlab software. Anxiety was assessed with State Anxiety Inventory（S- AI）and Trait Anxiety Inventory（T- AI）. Response time and inhibiting effects between two groups were compared by t test and Mann- Whitney U test ，respectively. Spearman correlation analysis was applied to determine the relationship between inhibiting effect and anxiety.【Results】The scores of S-AI and T-AI in POAG group were significantly higher than those in normal subjects（t = -3.912，P = 0.001；t = -2.127，P = 0.042）. There were statistically significant differences in the scores between groups even after gender was adjusted by using line regression model（β = 9.449，P = 0.000；β = 7.089，P = 0.018）. The average response time in POAG group（0.84±0.15）s was significantly longer than that in normal participants（0.75±0.14）s（t = -2.127，P = 0.039）. The difference was statistically insignificant when gender was adjusted by using line regression model（β = 0.063，P = 0.210）. The inhibiting effect in POAG group（0.126±0.089）s was longer than that in normal subjects（0.085±0.066）s（Z = -1.715，P = 0.086）. The inhabiting effect wasn′ t statistically significantly correlated with scores of S- AI and T- AI（P＞0.05）.【Conclusions】Patients with POAG showed severer anxiety than normal subjects but attention was comparable. There was no significant correlation between attention and anxiety.